Genome mining and molecular networking analyses have revealed a renewed capacity to uncover molecules and biosynthetic pathways from bacterial secondary metabolism. The present study describes the association of genome mining and molecular networking to correlate production of valinomycin cyclodepsipeptide derivatives by Streptomyces sp. CBMAI 2042. Additionally, expression of the entire gene cluster in a heterologous host successfully allowed the association of this biosynthetic assembly to the production of montanastatin, valinomycin and at least five structural analogues, revealing a plasticity of the modules and an interesting thioesterase activity capable of assembling two or three units of the tetradepsipeptide monomer.
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